16-nitro-alkyl steroids



United States Patent Patented Sept. 5, 1961 2,999,103 16-NITRO-ALKYL STEROIDS Richard Rausser, Union, and Eugene P. Oliveto, Glen Ridge, N.J., assignors to Schering Corporation, Binomfield, NJ., a corporation of New Jersey No Drawing. Filed May 1, 1959, Ser. No. 810,264 25 Claims. (Cl. 260-39145) This invention relates to novel steroids and to methods of preparing the same. More particularly the present invention is concerned with novel l-nitroalkyl pregnenes, and explicitly 16w(a-nitroalkyl)-4-pregnenes, the 9othal0 geno derivatives and 2l-esters thereof, and the corresponding 16w(a-nitroalkyl)-l,4-pregnadienes; and to the preparation thereof which includes reacting a steroid having the preguane carbon skeleton and a 16,17-double bond conjugated with a ketonic oxygen at the C position, with a nitroalkane and introducing a l7a-hydroxy group into the resultant nitroalkyl pregnane to cause the formation of a loot-(ot-nitroalkyl) -l7ot-hydroxypregnane which is or is convertible to the novel compounds of our invention as described more fully hereinafter. The terms, pregnane and pregnene as thus employed are intended to include both pregnenes and pregnadienes.

This application is a continuation-in-part of US. application Serial No. 733,843, filed May 8, 1958 by Richard Rausser and Eugene P. Oliveto, the inventors herein.

The novel pregnenes of our invention are represented by the following general formula:

CHzOR wherein [W W is CH -CI-I or CH=CH; Y represents keto-nic oxygen, (H, aOH) or (H, ,BOH); X is hydrogen or a halogen atom of an atomic weight less than 125, (Le. fluorine, chlorine or bromine) and wherein X is hydrogen when Y is (H, aOI-l); each of Z and Z is a member selected from the group consisting of hydrogen and an alkyl radical; and R is either hydrogen phosphate or acyl. Suitable acyl substituents at the C-21 position are carboxylic acyl radicals and preferably those containing from 1 to 8 carbon atoms such as alkanoates exemplified by acetate, propionate, cyclopentyl propionate, dimethylacetate, trimethylacetate, t-butylacetate, aryl carboxylates such as for example, phenoxyacetate, benzoate, sulfo-benzoate, phthalate; and heterocyclic carboxylates such as thiophene c'arboxylate, nicotinate, and the like.

Illustrative of the u-nitroalkyl substituents at the 16-carbon atom contemplated by our invention are 1606-(11lil'O- methyl), 16u-(a-nitroethyl), 16ot-(a-nitropropyl), 16ot-(unitro-2-methylethyl), 16a-(ot-nitrobuty1), 160L-(ot-llllllOlSO- butyl), loot-(wnitrodecyl), 16a-(a-nitrolauryl) and the like. The preferred 'a-nitroalkyl groups are the nitro-lower alkyls containing from 1 to 8 and most desirably l to 4 carbon atoms.

Certain of the novel compounds of our invention, that is, the 16a-(ot-nitroalky1)- substituted derivatives of contisone, hydrocortisone, prednisone, and prednisolone, nonhalogenated or halogenated in the 9wp0sition, as Well as their C21 esters (thus explicitly excluding the lla-hydroxy-16-nitroalkylated corticoids, which however, are valuable as intermediates in the formation of the corresponding 11-keto steroids as described herein), possess, as

has been noted above, valuable anti-inflammatory and diuretic (i.e. sodium and water excretory) properties. The nitroalkylated (notably the 16ot-nitromethyl-substituted) dienes with or without the 9ot-halogen substituent are particularly valuable therapeutic agents possessing significant therapeutic effects in the treatment of inflammatory diseases, such as for example, arthritis, dermatitis, asthma, and the like, and are normally administered in a daily maintenance dosage range of 0.50 to 10 mg. the preferred daily dosage of the non-halogenated dienes being from about 2 mg. to about 10 mg. and that of the halogenated, and preferably the 9ot-fluoro-alkylated dienes, being in the range of 0.5 mg. to 5. 0 mg. p

The nonahalogenated l6ot-(ot nitroalkyl) monoenes of our invention, in addition to being valuable intermediates in the formation of the corresponding dienes, are also active as anti-inflammatory agents and in inhibiting renal tubular reabsorption of sodium. A daily dosage of 5 mg. to 50 mg. is recommended for oral administration of these monoenes to induce sodium and water diuresis, e.g. in the treatment of ascities, congestive heart failure and cyclical edema.

Similarly, the Q-hflIOI6d(0L-IlltrOalkylatd) monoenes of our invention are also useful as anti-inflammatory agents, since they manifest an enhanced activity in the treatment of such inflammatory diseases as arthritis, dermatitis, asthma, and the like. When administered orally, the recommended daily dosage is in the range of 4 mg. to 15 mg.

When treatment so indicates, these 16nt-(ot-nitroalky1- ated) dienes and monoenes can be administered parenterally in the form of therapeutically acceptable solutions and suspensions, e.g., in aqueous media, or where oral administration is indicated, can be incorporated into tablets (normally from about 0.50 mg. to 5 mg. per tablet), eliXirs and other known pharmaceutical dosage forms by standard procedures. They can also be administered topically in the form of ointments or creams, or as solution, e.g. in dimethylacetamide or diethylacetamide, or in the form of suppositories dissolved or suspended in a fatty or waxy vehicle which melt at approximately body temperature, or as aerosols when mixed with suitable materials, such as isopropyl myristate and dichloro difluoromethane (Freon).

it has been known heretofore that the substitution of an tit-halogen atom for hydrogen at the C-9 position will enhance the anti inflammatory properties of the corresponding parent steroid, e.g. prednisone, prednisolone, cortisone, hydrocortisone. Unfortunately, a concomitant and significant increase in the salt retaining properties of such 9ot-halogenated derivatives occurs. It is worthy of particular note, therefor, that the 9ot-halogeno-l6-(u-nitroalkyl) compounds of the present invention effect the retention of the enhanced anti-inflammatory activity afforded by the presence of the 9ot-halogen while inhibiting the anticipated increase in salt retention above referred to. Similarly the salt retaining properties of the non-halogenated steroids such as for example, cortisone of the present invention can also be employed in the preparation of the loa-alkyl and 16,8-alkyl steroids of that application. The reaction sequence for the preparation of l6m-methyland 16fl-methyl steroids from the corresponding 16a-nitromethyl steroids is as follows. The l6a-nitromethyl derivative is reduced, eg. with tin and hydrochloric acid, to the l6a-aminomethyl steroid, which, in turn, is quatern'med with methyl iodide and subsequently pyrolyzed to form the corresponding l6-methylene derivative, which, in turn, converts to the l6-metl1yl-20-ketol6(l7)-dehydro compound. Peroxidation of this latter product with hydrogen peroxide will cause the formation of the 160:,17oz-OXidO, 16/3-methyl-20-keto steroid which when reacted with hydrobromic acid, for example, forms the intermediate bromohydrin which rapidly dehydrobrominates and is then reduced to yield a mixture (separable by column chromatography) of 16a-methyl and l6B-methyl derivatives of, for example, 4-pregnenel7a,21-diol-3,l1,20-trione, 4-pregnene llti,l7ot,2l triol- 3,20-dione and their 9a-halo (cg. fluoro) derivatives. These monoenes can be converted to the corresponding 1,4-dienes microbiologically employing, for example, the microorganism Bacillus splmei'icus (A.T.C.C. 7055) or Corynebaczerium simplex (A.T.C.C. 6946). These reaction steps are carried out by procedures Well known to steroid chemists. It will, for example, be evident that, should higher homologues of nitromethane be employed {c.g. nitroethane, 2-nitropropane, nitrobutane and the like) on a IG-dehydrosteroid such as 3a-acetoxy-l6-pregnene-ll,20-dione and the resultant product taken through the procedural steps described above, the corresponding alkyl homologues will result in the C-l6 position (eg. 1 6-ethyl-16-dehydro-ZO-ketosteroids, l6-isopropyll 6-dehydro-ZO-ketosteroids etc). The l6-alkyl-l6-dehydro-20- ketosteroids resulting from the pyrolysis of the quaternized steroids can be converted to corticoids in the manner already described, e.g., hydrogenation of the double bond with palladium on carbon or Raney nickel, introduction of the l7-hydroxy group by means of enolacylation, preferably enolacetylation and peroxidation (cg. acetic anhydride and p-toluenesulfonic acid followed by treatment with peracetic acid and sequential treatment with dilute alkali (eg. N aOH)), introduction of the 21- acetate by means of bromination and acetoxylation (sodium acetate in dimethylformarnide) and elaboration of the A-ring in the conventional manner to introduce the A*- or A -double bond(s) and 3-keto group. Alternatively, they may be epoxidized with alkaline hydrogen peroxide to yield l6a,17ct-epoxy, l6B-alkyl-20-keto steroids. Those are then treated with a hydrohalic acid such as hydrogen bromide and reduced, e.g. with Raney nic tel or palladium and hydrogen. This results in the formation of both 16u-alkyl-l7e-hydroxy-20-keto steroids, and l6,8-alkyl-17a-hydroxy-20-keto steroids. After separation, e.g., by column chromatography, these are then handled in the usual fashion, e.g., the ZI-acetate is introduced via bromination and acetoxylation, and the A-ring elaborated via oxidation of the 3-hydroxyl, followed by bromination-dehydrobromination to give an unsaturated 3-ketone.

The lfia-(a-nitro-lower alkyl) compounds of our invention are desirably prepared by the procedure, wherein a 3-oxygenated 20-keto-l6-pregnene such as illustratively, 3a-hydroxy-16pregnene-l1,20-dione, is reacted with a lower nitroalkane, such as for example, nitromethane, in the presence of a strong base to effect the production of the corresponding saturated l6a-(nitromethyl) derivative, a 3oxygenatcd 20-keto 16a (a-nitromethyl)-pregnane, e.g., 3ot-hydroxy-l6u-(ot-nitromethyl) pregnane-1l,20-dione. The nitromethane is employed merely for purposes of illustration, but it will be apparent that where it is desired to prepare other l6a-(a-nitro-lov1er alkyl) compounds such as the corresponding l6a-(a-nitroethyl), l6u-(a-nitropropyl) and l6a-(a-nitro-a-methylethyl) substitutcd steroids, nitroethane, l-nitropropaneand Z-nitropropane respectively are employed.

The product of this nitromethylation reaction, 3a-hydroxy-l6a-(nitromethyl)-pregnane-11,20-dione, is then hydroxylated by standard procedures at the C-l7 position, as for example, by enolacylation, preferably enolacetylation, and peroxidation e.g. with acetic anhydride and p-toluenesulfonic acid, followed by reaction with peracetic acid and then by alkaline hydrolysis to form 301,170- dihydroxy-l6a-(nitromethyl) -pregnane-1 1,20-dione. Introduction of the C-21 acetoxy group, or other ester thereof, is accomplished by known methods, such as for example, by bromination of the 0-21 methyl group in an inert solvent such as chloroform with the sequential reaction of this product with sodium or potassium acetate in acetone or dimethylformamide. The product thus formed is 3a,l7a,21-trihydroxy-16a-(nitromethyl)-pregnane-1l,20-dione ZI-acetate. The hydroxyl group at position 3 in this latter compound is then transformed to a keto group, preferably by means of N-bromoacetamidc to cause the production of l7a,21-dihydroxy-l6a-(nitromethyl) pregnane 3,11,20 trione Zl-acetate. Other known equivalent oxidizing agents such as N-bromosuccinimide, chromium trioxide -pyridine, chromium trioxideacetone-sulfuric acid can, of course, also be employed. The A -double bonds are then introduced into the A-ring by dihalogenation, c.g. dibromination of this 3-keto dcrivative by the rapid introduction of bromine in a suitable non-reactive organic solvent such as dioxane followed by dehydrohalogenation in a conventional manner such as by refluxing the brominated steroid with dimethylformamide in the presence of calcium or lithium carbonate and lithium chloride or collidine or the like. The resultant product is l6rt-(nitromethyl)-prednisone Zl-acctate.

Also, the l7a,2l-dihydroxy-l6e'(nitromethj/U-prcg nane-3,ll,20-trione 2l-acetate intermediate obtained by the procedure described above, can be modified by introduction solely of the A -double bond to form (nitromethyD-cortisone acetate. This is normally accomplished by halogenation of the former compound (preferably with bromine in a medium such as tert.-butyl alcohol), at about 30 C. to 35 C. with subsequent dehydrohalogenation by conventional reaction, as with semicarbazide followed by hydrolysis of the resultant 3- mono-semicarbazone with pyruvic acid or the like.

The loa-(nitromethyl)-cortisone acetate so formed or the corresponding diene, for example, 16a-(nitromcthyl)- prednisone Ill-acetate can then be saponitied by reaction with conventional hydrolytic reagents such as sodium or potassium bicarbonate, sodium hydroxide, sodium methoxide or p-toluenesulfonic acid to remove the acyl radical at C-21 and form the alcohol, l6a-(nitromethyl)- cortisone, or l6e-(nitromethyl)-prednisone. These compounds can be re-esterified if desired to further enhance the duration of activity, by formation of the propionate, t-butylacetate, trimethylacetate, succinate, phosphate, 0- sulfobenzoate and the like.

The l6a-(a-nitro-lower alkyl) substituted cortisone acy-lates such as the l6a-(wnitromethyl)-cortisone acetate can also be converted to l6a-(a-nitromethyl)-hydrocortisone by reaction thereof initially with semicarbazidc, for example, to produce the lfist-(whitesmethyll'coriisone acetate 3,20-bis-semicarbazone, which is reacted with a metal borohydride reducing agent such as sodium or potassium borohydride to reduce the 0-11 keto group to the llfl-hydroxy group while simultaneously saponifying the C-12 position to form 16a-(nitromethyl)-hydrocortisone-3,20-bis-semicarbazone. The latter product is then hydrolyzed by conventional procedures, e.g. dilute aqueous hydrochloric acid or pyruvic acid, to form 16a- (nitromethyl)-hydrocortisone. This latter intermediate can then be transformed microbiologically according to procedures analogous to those descibed in Belgian Patent No. 540,478 employing a dehydrogenating microorganism, e.g. Corynebacterium simplex (A.T.C.C. 6946), Bacillus sphaericus (A.T.C.C. 7055), to form the valuable anti-inflammatory steroid, l6a-(nitromethyl)-prednisolone. The compound can then in turn be re-esterified, as for example, to form an acylate such as la-(nitromethyl)-prednisolone 21-acetate by reaction thereof with acetic anhydride in a suitable non-reactive organic solvent such as pyridine or lutidine and subsequently crystallized out.

In one preferred method the 9a-halogen atom, and preferably the 9a-fiuorine atom, is introduced into the 16ot-(nitromethyl) monoene and diene 21-esters as represented, for example, by 16u-(nitromethyl)-hydrocortisone acetate and l6ot-(nitromethyl)-predniso1one acetate respectively, by first dehydrating 16a-(nitromethyl)- prednisolone acetate, for example, with an alkyl or aryl sulfonyl chloride, e.g., methanesulfonyl chloride, in an alkaline organic media, for example, pyridine, to effect the production of the corresponding M -derivative, e. g. 16a-(nitromethyl) -A -pregnatriene-170:,21-di0l- 3,20-dione 2l-acetate and in the case of the monoene, 16w (nitromethyl) N pregnadiene 171:,21- diol-3,20-dione 21-acetate. Placement of a halogen, and preferably bromine at this point in the procedure at the 9a-carbon position is accomplished. by conventional means, such as, for example, by reacting the A steroid with hypobromous acid which can be and indeed is preferably prepared in situ from the reaction of for example N-bromoacetamide and perchloric acid yielding directly the corresponding 9a-bromo-16ot-(nitromethyl)- prednisolone 21-acetate, and 9oc-bl0II10-l6ct-(HltI'OII16thyl)-hydrocor tisone 2l-acetate. Esters of these types are useful anti-inflammatory compounds and are at the same time readily converted by oxidation to the corresponding ketones by known techniques such as, for example, with chromium trioxide in pyridine and readily hydrolyzed at the C-21 position by such standard hydrolytic agents as aqueous methanolic potassium bicarbonate, sodium carbonate, concentrated hydrochloric acid in methanolchloroform or like substances as disclosed hereinabove, to prepare the corresponding 2l-alcohols.

A further method of introducing a 9a-halogen atom into the 16vt-nitro monoenes and dienes of our invention utilizes as starting compounds the 9ot-bromohydrins prepared as above, which can be refluxed with mild alkali, such as illustratively, sodium acetate in methanol, to form the corresponding 9,8,11,8-oxido derivatives, 93, 11,8 oxido 16a (nitromethyl) A pregnadienel7a,21-diol-3,20-dione 21-acetate, 9,B,llfi-oxido-l6ot-(nitromethyDM-pregnene-17,2l-diol-Zl-acetate. These latter compounds are then reacted with hydrogen fluoride in chloroform with or without ethanol and/or tetrahydrofuran to cause the formation of 9ot-fill01O-l6a-(I1i'tl0- methyl)-prednisolone acetate and 9OL-flllOI'O-160t-(Illtl'0- methyl)-hydrocortisone acetate. Simularly the substitution of anhydrous hydrogen chloride for hydrogen fluoride in this reaction results in the production of the corresponding 9oc-Chl01'0 derivatives. Again, these compounds can be oxidized to the corresponding ll-ketones by standard agents, e.g. chromium trioxide or N-halosucoinimides, or hydrolyzed to the corresponding 21-alcohols by standard saponification procedures (e.g. aqueous methanolic potassium bicarbonate or concentrated hydrochloric acid in methanol). Thus the preferred halogenated compounds of our invention, 9u-fluoro-l6a-(unitro-lower alkyl)-prednisolones and 90t-flll0l'O-l60t-(Otnitro-lower alkyl)prednisones and particularly 9a-fluoro- 16a-(nitromethyl)-prednisolone and 9ot-fit101'0-16a-(Ditt0- methyl) -prednisone are prepared by these procedures.

Thus far, we have described as starting materials only compounds which are available from bile acids. There are, however, several other classes of starting material which are extremely useful, e .g. diosgenin and smilagenin. For example, diosgenin is readily converted to 16-dehydropregnenolone which can be converted readily to 160:- (nitromethyl)-pregnenolone by the addition of nitromethane. This compound is selectively hydrogenated, e.g. by means of palladium catalyst, to Mo -(nitromethyl)-a1lopregnane-3B-ol-20-one and the cortical side chain elaborated in the usual way (enolacetylation at C-17 and epoxidation to give the 17a-hydroxy-20-ketone followed by bromination and acetoxylation at C-21). The

C-3 hydroxyl is oxidized to the ketone, a 2,4-dibromide is prepared using bromine and dioxane or other inert solvent and dehydrobromination is effected by means of collidine or dirnethylformamide to yield 16a-(nitromethyl)-A -pregnadiene-17a,21-diol-3,20 dione 21-acetate. This can be hydroxylated at position (3-11 to yield either l6ot-(nitromethyl)-prednisolone or l6a-(nitromethyD-ll-epiprednisolone. This last compound may undergo acyloxylation, e.g., acetylation selectively at C-21 to yield a mono acetate with sequential oxidation at C-ll to give 161x-(nitromethyl)-prednisone ZI-acetate. Also, loa-(nitromethyb-ll-epi-prednisolone 21-acetate can be dehydrated at C-ll by tosylation of the C-llahydroxyl followed by treatment with sodium acetate to yiled 16a-(nitromethyl)-A pregnatriene 17u,21- diol-3,20-dione 21-aceta-te, a compound previously described as an intermediate for the preparation of 9a-halo- 1611- nitromethyl) -corticoids.

Other important starting materials are the l2-oxygenated sapogenins, such as hecogenin, rockogenin and gentrogenin. For example, hecogenin has been converted to 1l-keto-l6-dehydro-allopregnane-BB-ol-20-one. A lfia-(nitrornethyl)-substituent can be introduced by means of the standard addition of nitromethane and the side chain and A-ring elaborated as described above in the usual manner to contain the A or A -double bond(s) and 3-keto group.

The following examples are further illustrative of the invention.

EXAMPLE 1 16a(Nitromethyl) cortisone Z1 -acetate A. lfia-(NITROMETHYL) PREGNANE-3a-OL-11,20- IDIONE To a solution of 2.0 g. of 16-pregnene-3a-ol-1 1,20- dione 3-acetate in 50 ml. of t-butyl alcohol containing about 1 gram of potassium t-butylate there is added 3 ml. of nitromethane. The reaction mixture is allowed to stand for 24' hours at room temperature, and then is poured into a large volume of water. The resulting precipitate is filtered, air-dried, and crystallized from acetone-hexane to give 1.3 g. of 16a(nitromethyl) pregnane- 3u-ol-11,20-dione.

B. 16a-(NITROMETHYL) PREGNANE-3a,17a DIOL-11,20

DIONE A solution of 3.4 g. of 16ot-(nitromethyl)pregnanc- 3a-ol-ll,20-dione prepared as in above Example 1A, in 76 ml. of acetic anhydride containing 1.9 g. of p-toluenesulfonic acid is kept at C. for six hours; during which time about 8 ml. of distillate is removed every half hour by the application of vacuum. The resulting oily residue is dissolved in 50 of benzene and Washed three times with water, then with a solution of 1.0 g. of sodium acetate in 10 ml. of water. The benzene layer is dried over magnesium sulfate, filtered, and then stirred for 18 hours at 25 C. with a mixture of 0.26 g. of sodium acetate in 6 ml. of commercial 40% peracetic acid. Any excess peracetic acid is then destroyed by the dropwise addition of a solution of 8 g. of sodium sulfite in 25 ml. of water, while maintaining the temperature between 10-20" C. An additional 0.8 g. of sodium sulfite is then added, and the mixture stirred overnight until a starch-iodide test is negative. The benzene layer is separated, Washed three times with water, and evaporated, To the resulting residue which is dissolved in ml. of methanol, there is added a solution of 1.8 g. of sodium hydroxide in 20 ml. of water, and the mixture refluxed for 15 minutes. After neutralization with 2 ml. of glacial acetic acid, the solution is concentrated under reduced pressure to a volume of about 40 ml. This concentrate is poured into a mixture of ice and water. The resulting precipitate is filtered and dried, yielding 3,3 g. of a solid which is chromato I graphed on Florisil. The material eluted with 33-50% ether-hexane is combined and crystallized from acetonehexane to yield 1.1 g. of l6a-(nitromethyl) pregnanc- 30L,17lZ-di0i-11,20-dl0fl6.

C. lfia-(NITROMETHYL) PREGNANE-3a,17a(2l'lRl0L- 11,20-DIONE 2l-ACETATE A solution of 181 mg. of 1604-(I1it1'0ffi6i'hY1) pregnanc- 3a,]7ocdi0l-ll,20di0lte, prepared as in Example 1B, in 9 ml. of Cl. chloroform (containing a few drops of chloroform previously saturated with hydrogen bromide) is brominated at -25 C. by the addition (over a three hour period) of 83 mg. of bromine in 5 ml. of chloroform. The solvent is distilled in vacuo then 5 ml. of dirnethylformamide and 0.5 gram of sodium acetate are added. The mixture is stirred at 60 C. for four hours, then poured into water, and the resulting precipitate is filtered and dried to yield 180 mg. of a solid which on recrystallization from acetone-hexane give 120 mg. of 16a (nitromethyl) pregnane-3a,17a,2ltriol-11,20-dione 2l-acetate.

D. IGa-(NITROMETHYL) PREGNANE-17a,2l-DIOL-3,11,

QO-TRION'E ZLACETATE A solution of 90 mg. of l6a(nitro:methyl) pregnanc- 3a,l7a,2l-triol-l1,20-dione 21-acetate (the product of Example lC) in ml. of 80% acetone-water is cooled to 10 C. One drop of concentratde hydrochloric acid is added, along with 75 mg. of N-bromos uccinimide, and the mixture is ltept in the dark for 22 hours at 10 C. Excess sodium sulfite solution is then added, and the mixture concentrated in vacuo to a small volume from which precipitates 60 mg. of a solid which is crystallized from acetone-hexane to give 40 mg. of 16a(nitromethyl) preguane-17a,2l-diol-3,11,20-trione ZI-acetate. E. 4B-BROMO-16a.-(NITROMETHYL) PREGNANE-17a,21-

DIOL-3,11,20-TRIONE 21-ACETATE A solution of 84 mg. of l6a-(nitromethyl) pregnanc- 17a,21-ii0l-3,i1,20-trl0ne 21-acetate in 3 ml. of t-butyl alcohol and 3 ml. of methylene chloride is brorninated at 30-35 C. by the rapid addition of a solution of 32 mg. of bromine in 3 m1. of t-butyl alcohol. After bromination is complete as evidenced by the discharge of color (about 1 /2 hours), the solution is evaporated in vacuo to a residue. This residue is slurried with water, filtered and dried to yield 90 mg. of a solid substantially of 4,8- bromo l6a-(nitromethyl) pregnanc-17a,21-diol-3,l1,20- trione ZI-acetate of which is used without further purification in the conversion described in the procedure immediately following.

F. lfia-(NITROMETHYL) CORTISONE ZI-ACETATE The 4fi-bromo-IGm-(nitromethyI) pregnane-17a,21-diol- 3,11,20-trione 21-acetate'of above Example 1B is dissolved in a mixture of 30 mg. of semicarbazide, 4 ml. of t-butyl alcohol and 2 ml. of methylene chloride, and the solution stirred in a nitrogen atmosphere for two hours at 25-30 C. The solvent is distilled in vacuo and the resulting residue dissolved in 5 ml. of 80% acetic acidwater along with excess 70% pyruvic acid. After standing at room temperature for hours, the reaction mixture is brought to about pH 7 with dilute (5%) sodium hydroxide. and then is extracted with methylene chloride. The organic extracts are combined and evaporated to a residue, which is chromatographed on Florisil. The crystalline material mg.) obtained from ether-hexane and 100% ether eluates, is crystallized from acetonehexane to yield 20 mg. of l6u-(nitromethyl) cortisone 2l-acetate,

M32 238 III. (e 14,500)

EXAMPLE 2 16u-(nitromethyl) cortisone One gram of l6a-(nitromethyl) cortisone 21-acetate (prepared as in Example IE) is dissolved in 25 ml. of

methanol and 5 m1. of water containing 0.2 g. of potassium bicarbonate. This solution is refluxed for /2 hour, then concentrated in vacuo to a residue. Water is added to this residue, and the resulting precipitate is filtered, dried, and crystallized from acetone-hexane to give 16a- (nitromethyl-cortisone,

A532 238 my (5 14,200) EXAMPLE 3 l6a-(Nitromethyl) hydrocortisone A. IGa-(NITROMETHYL) CORTISONE 3,20-BIS-SEMI- CARBAZONE A mixture of 3.0 g. of 16a-(nitromethyl) cortisone, 5.0 g. of semicarbazide hydrochloride, 4 g. of pyridine, 30 ml. of water and 140 ml. of methanol is refluxed for 16 hours. The solution is then concentrated to about 30 ml. and poured into water to precipitate a solid substantially of la-(nitromethyl) cortisone-3,20-bis-semicarbazone which is used without further purification in the reaction immediately following.

B. IGa-(NITROMETHYL) HYDROCORTISONE 3,20-

BIS-SEMICARBAZONE A solution of 3.0 g. of the bis-semicarbazone of above Example 3A and 2 g. of potassium borohydride in ml. of tetrahydrofuran and 50 ml. of water is refluxed for 6 hours. The solution is cooled, and acetic acid added until the solution is approximately pH 5.5. The organic solvent is distilled from the reaction mixture and the resulting solid precipitate in the aqueous residue is filtered to give loa-(nitromethyl) hydrocortisone 3,20-bis-semicarbazone of sufficient purity to be used directly in the following reaction.

C. lilo-(NITROMETHYL) HYDROCORTISONE EXAMPLE 4 16a-(nitr0melhyl) hydrocortisone 21 -acetate To 0.5 g. of 16a-(nitromethyl) hydrocortisone, prepared as in Example 3, there is added 0.3 ml. of acetic anhydride in 2.3 ml. of pyridine. After standing for one hour at room temperature, the mixture is poured into ice and hydrochloric acid. The resulting precipitate is filtered and recrystallized from aqueous methanol to yield l6a-(nitromethyl) hydrocortisone Zl-acetate.

EXAMPLE 5 9a-br0m0-16u-(nitromethyl) hydrocorlisone 21 -ucetate A. ltict- (NITROMETHYL) -4,9PREGNADIENE-17a,21-

DIOL-3,20-DIONE ZI-ACETATE To a solution of 16a-(nitromethyl) hydrocortisone 2l-acetate (0.3 g.), prepared as in Example 4, in 5 ml. of pyridine there is added 0.2 ml. of benzenesulfonyl chloride in 3 ml. of pyridine. The solution is allowed to stand for 4 hours, then is poured into ice-hydrochloric acid. A solid precipitates which is filtered and crystallized from acetone-hexane to give l6a-(nitromethyl)-4,9-pregnadiene-l7a,21-diol-3,20-dione ZI-acetate,

A 239 III .1. (6 15,500)

men

B; BIZ-BROMO-l Sa- (N ITROMETHYL) HYDROCORTISONE '21-ACETATE A suspension of 0.2 g. of "the -16a (nitroniethyl)-'4,9- pregnadiene of above Example 5A in 20 ml. of purified dioxane, 2 ml. of water and containing 0.1 gofN-bromoacetamide and 1 ml. of 1.5 N perchloric acid is gently agitated for two hours. During this time the mixture becomes homogeneous. A solution of 0.2 g. of sodium sul-fite in 2 ml. of water is then added, and the'solution extracted with methylene chloride. The organic extracts are washed with water, dried and evaporated to a solid residue which is crystallized from acetone to give S ot-brorno-l6a-(nitromethyl) hydrocor'tisone 21-acetate,

A2552 243 m (6 15,700) EXAMPLE '6 9a-brom0-l6u-(nitromethyl) hydrocortisone A mixture of 0.3 g. of 9a-bromo-l'fiu-(nitromethyl) hydrocortisone 21-acetate, prepared as in Example 5, in 50 ml. of methanol, 10 ml. of chloroform, 3 ml. of water and 3 ml. of concentrated hydrochloric acid is allowed to stand 48 hours at room temperature. Water is then added, and the mixture extracted with methylene chloride. The organic extracts are washed with water, dried and concentrated to a residue. Crystallization of this residue from acetone yields 9a-bromo-l6ot-(nitromethyl) hydrocortisone.

EXAMPLE 7 9ot-flu0r0-]6a-(nitromethyl) hydrocortis'one 21-acetate A. ssnia-oxrno-lsa-(NIT OMETHYLytPREGNENE- 17a,21'DIOL-3,20=DIONE 21-ACETATE To 9a-bromo16u-(nitromethyl) hydrocortisone 21- acetate (0.3 g.), prepared as in Example 5, in 20 ml. of methanol there is added 0.3g. of potassium acetate. The mixture is refluxed for 2 hours, then concentrated in vacuo to a residue. Water is added to the residue, and a solid separates which is filtered and crystallized from methanol-water to give 0.1 g. of 9/3,11fi-oxido-16u-(nitromethyl)-4-pregnene-l7a,21-diol-3,20-dione 2l-acetate.

B. Qa-FLUORO-lfia-(NITROMEQJHYL) HYDROCORTI- SONE 21-ACETATE A solution of 0.1 g. of the 9p,llfl-oxido=4 -'p'r'egnene of above Example 7A in 5 ml. of alcohol-free chloroform is saturated with anhydrous hydrogen fluoride at C. The mixture is allowed to stand 4 hours at 0 C., then concentrated to a residue in vacuo, which when crystallized from acetone-hexane gives 50 mg; of 9ot fluoro=l6u- (nitro methyl) hydrocortisone ZI-acetate.

EXAMPLE 8 9a-flu0r0-16a (nitromethyl) hydrocortison'e 9oa-fluoro-16a-(nitromethyl) hydrocortisone 21-acetate, prepared as in Example 7, is hydrolyzed with hydrochloric acid in chloroform-water in the manner described in Example 6 to give 9a-fluoro-16ot-(nitromethyl) hydrocortisone. U H

EXAMPLE 9 Qua-Ch Zora-1 6 (Z- nitromethyl) hydrocor'tison'e -21 acetate A solution of 0.3 g. of the 95,11fl-oxido-4-pregnene of Example 7A in 40 ml. of alcohol-free chloroform is saturated at 0 C. with anhydrous hydrogen chloride, and the mixture allowed to stand at 0 C. for six hours. The solvent is distilled in vacuo from the reaction mixture leaving a residue which, when crystallized from acetone-water, gives 0.2 g. of 9oc-chloro-16a-(nitromethylfhydrocortisone, Am 241 m -(e 15,900) EXAMPLE -10 9u-chl0r0-1 6 a-(nitromethyl) hydrocortisone In the manner described in Example 6, 9u chloro-16anitromethyl) hydrocortisone ZI-acetate, .prepared as in Example 9, is converted to Qa-chloro-l6a-(nitromethyl) hydrocortisone by means of hydrochloric acid in methanol-chloroform-Water.

EXAMPLE 11 1 6ot-(nitromethyl) prednisone ZJ-acetate A. ZA-DIBROMOJGa-(NITROMETHYL) PREIGNANE- 17 a,21-DIOL-3,11,20-TRIONE QLACETATE A solution of 180 mg. of loot-(nitromethyl) pregnanc- 17ot,2l-diol-3,11,20-trione 21-acetate, the compound of Example 1D, in 3 ml. of dioxane is dibrominated in positions 2 and 4 by the rapid addition of 130 mg. of bromine in 1 ml. of dioxane at room temperature. The solution is poured into water and the precipitated solid is filtered, yielding 190 mg. of 2,4-dibromo-16a-(nitromethyl) pregnane-17a,21-diol-3,11,20-trione 21-acetate of sufficient purity to be used in the conversion described in the following procedure.

B. 16a-(NITROMETHYL) PREDNISONE 21-ACETA1E The 2,4-dibromide (190 mg.) prepared as in above Example 11A, is dehydrobrominated by refluxing for 2 hours with 4 ml. of dimethylformamide containing 30 mg. of lithium carbonate and 30 mg. of lithium bromide. The mixture is poured into dilute hydrochloric acid and extracted with methylene chloride. The organic extract is evaporated to a residue (155 mg), which is chromatographed on Florisil. The fractions obtained by elution with 20% ether-hexane are crystallized from acetonehexane to give mg. of l6a-(nitr0methyl) prednisone 21-acetate,

AEQQ 237 m (6 14,600)

EXAMPLE 12 1606- (nitromethyl) prednisone 16ot-(nitromethyl) prednisone 2l-acetate (0.5 g.), the compound of Example 11, when hydrolyzed by means of aqueous alcoholic potassium bicarbonate in the manner described in Example 2, yields ld-(nitromethyl) prednisone.

A further method of preparation of the compound of this example is as follows:

Bacillus sphaericus var. fusiformis (A.T.C.C. 7055) is incubated on a nutrient agar (composed of Bacto-beef extract, 3 g., Bacto-peptone, 5 g.; sodium chloride, 8 g.; agar, 15 g.; tap water, 1 liter) for 24 hours at 28 C.

To ml. of a sterile nutrient broth (composed of Bacto-beef extract, 3 g.; Bacto-peptone, 5 g.; per liter of tap water) in a 300 ml. flask is added one loopful of the incubated culture and the broth mixture is further incubated for 24 hours at 28 C. on a shakmg machine. The broth culture so obtained is employed as an inoculum (1%).

Into each of ten flasks containing 100 ml. of sterile nutrient broth is added 1 m1. of the inoculum. The flasks are agitated on a rotary shaker for 8 hours at 28 C. and 240 strokes per minute. After this growth period, a solution of 25 mg. of 16ot-(nitromethyl) cortisone (the compound of Example 2, in 0.5 mL-of methanol is aseptically added to each flask which in turn is reshaken and incubated for an additional 24 hours. The final pHis 7.8. I

The contents of the flasks are then combined and extracted three times with 2 liters of chloroform per extraction. The combined chloroform extracts are evaporated to dryness yielding 310 mg. of crude product. The crude steroid is purified by chromatography on a chromatographic system described by G. M. Shull, Abstracts of Papers of the 126th Meeting of the American Chemical Society, December 12-17, 1954, page 911, paper No. 24. Chromatographic evaluation shows a quantitative conversion of the starting material to the diene when an auth'entic sample of the lou-(nitromethyl) prednisone isu's'ed as a control.

Alternatively, the crude product is recrystallized from acetone affording 210 mg. of 16a-(nitromethyl) prednisone.

EXAMPLE 13 1 6 a-(nitromethyl prednisolone l6a-(nitromethyl) hydrocortisone, the compound of Example 3, is fermented by means of Bacillus sphaericus var. fusiformis (A.T.C.C. 7055) in the manner described in the alternate procedure of Example 12 to produce 16a-(nitromethyl) prednisolone.

EXAMPLE 14 I 6ot-(nitromcthyl) prednisolone ZI-acetate lfia-(nitromethyl) prednisolone of Example 13, is treated with acetic acid and pyridine in the manner described in Example 4 to yield l6u-(nitromethyl) prednisolone 2l-acetate.

EXAMPLE 15 9m-br0mo-16a-(nitromethyl) prednisolone Z1 -acetate :1. 16a(NITROMETHYL)-1,4-9(11)-PREGNATRIENE- 17a,21-DIOL-3,20-DIONE 2'1-ACETATE m 228 my (e 15,100)

max.

13. 9a-BROMO-16a-(NITROMETHYL) PREDNISOLONE 21- ACETATE A suspension of 0.3 g. of the 160L-(Hllll'OIIl6thYl) pregnatriene of above Example 15A in 30 ml. of purified dioxane to which has been added 3 ml. of water containing 0.15 g. of N-bromoacetamide and 1.5 ml. of 1.5 N perchloric acid, is gently agitated for a period of two hours, during which time the mixture becomes homogeneous. A solution of 0.3 g. of sodium sulfite in 3 ml. of water is then added, and the reaction mixture is extracted with methylene chloride. The organic extracts are washed with water, dried over magnesium sulfate, filtered and evaporated. The resulting solid is crystallized from acetone to yield 0.2 g. of 9a-bromo-l6a-(nitromethyl) prednisolone 21-acetate,

A3 3 242 m (e 15,200

EXAMPLE 16 Qa-BROMO-lfia-(NITROMETHYL) PREDNISOLONE A gram of 9a-bromo-l6a-(nitromethyl)-prednisolone 2l-acetate prepared as in Example is hydrolyzed with hydrochloric acid in chloroform-methanol-water in the manner described in Example 6 to give 90L-b1'OII10-16IX- (nitromethyl) prednisolone.

EXAMPLE 17 9oc-flu0r0-16a-(nitromethyl) prednisolone 21-aceta1e A. QBJJEOXIDO-lfia- NITROMETHYL) -1,4-PREGNA- DIENE-1Ta,21-DIOL'3,20-DIONE 21-ACETATE 12 ,B. sa-r'nnono-ieanrrnormrnrn) PREDNISOLONE zrncnrarn A solution of 0.2 g. of the 9B,115-oxido-1,4-pregnadiene of Example 17A in 20 ml. of alcohol-free chloroform is saturated with anhydrous hydrogen fluoride at 0 C. The mixture is allowed to stand 4 hours at 0 C., then concentrated to a residue under reduced pressure. Crystallization of the residue from acetone-hexane gives 0.1 g. 9a-fluoro-16a-(nitr0methyl) prednisolone 21-acetate,

max.

238 m (6 15,250) EXAMPLE 18 9a-flu0r0-16a-(nitromethyl) prednisolone 9a-fluoro-16u-(nitromethyl) prednisolone 21-acetate, the compound of Example 17, is converted to 911-1111010- 16,8methylprednisolone by means of hydrochloric acid in methanol-chloroform-water in the manner described in Example 6.

EXAMPLE 19 9u-chl0r0-16a-(nitromethyl) prednisolone 21-acetate A solution of 0.2 g. of 9,6,11,8-oxido IGa-(nitromethy-l) 1,4-pregnadiene-17a,21-dio1-3,20-dione 2l-ace tate, the compound of Example 15A, in 30 ml. of a1cohol-free chloroform is saturated at 0 C. with anhydrous hydrogen chloride, and the mixture allowed to stand at 0 C. for 6 hours. The mixture is concentrated under reduced pressure to a residue which is crystallized from acetone-water to give 0.15 g. of 9a-Ch1OI'0-16a(flitr0- methyl) prednisolone 21-acetate,

Am 240 m (6 14,900)

EXAMPLE 20 Qa-chloro-I6a-(nitromethyl) prednisolone In the manner of Example 6, 9a-chloro-16u-(nitro methyl) prednisolone 2l-acetate is converted to 9a-chloro- 16a-(nitromethyl) prednisolone.

EXAMPLE 21 9w uor0-16a-(nitr0methyl) prednisone ZZ-acerate To a solution of 0.2 g. of 9a-fiuoro-16a-(nitromethyl) prednisolone ZI-acetate, the compounds of Example 17, in 10 ml. of acetic acid is added dropwise a solution of 40 mg. of chromium trioxide in 1 ml. of water and 3 ml. of acetic acid. The resulting mixture is allowed to stand 5 hours, then diluted with water and extracted with methylene chloride. The organic extracts are washed with water, dried over magnesuim sulfate, filtered and evaporated leaving a residue which is crystallized from methanol to give 0.1 g. of 9a-fluoro-l6a-(nitromethyl) prednisone 21-acetate.

EXAMPLE 22 9a-flu0r0-16u-(nitromethyl) prednisone In the manner described in Example 6, Qa-flUOrO-lfia- (nitromethyl) prednisone ZI-acetate, the compound of Example 21, is hydrolyzed to 9a-fiuoro-l6a-(nitromethyl) prednisone by means of HCl in methanol-chloroform water.

EXAMPLE 23 9a-chloro-16a-(nitr0methyl) prednisone .A. Qa-CHLORO lGa-(NITROM'ETHYL) PREDNISONE 21-ACETATE 9ot-chloro-l6oc-(nitromethyl) prednisolone ZI-acetate,

the compound of Example 19, is reacted with chromium a trioxide in aqueous acetic acid in the manner of Example 21 to give 9a-chloro-16u-(nitromethyl) prednisone 21- acetate.

B. Qa-CHLORO-lfia-(NITROMETHYL) PREDNISONE In the manner described in Example 6, the Zl-acetate 13 ester of Example 23A is reacted with hydrochloric acid in methanol-chloroform water to give 9otchloro-l6a-nitromethyl) prednisone.

EXAMPLE 24 9u-br0mo-16a-(nitr0methyl) prea'nisone A. QKZ-BROMO-lfid-(NITROMETHYL) PREDNISONE ZI-ACETATE 9a-bromo-l6u-(nitromethyl) prednisolone 21-acetate, the compound of Example 15, is reacted with chromium trioxide in aqueous acetic acid in the manner of Example 21 to give 9a-bromo-l6u-(nitromethyl) prednisone 21- acetate. i

B. 9a-BROMO-16a(NITROMETHYL) PREDNISONE In the manner of Example 6, the 21-acetate ester of Example 24A, is reacted with hydrochloric acid in methanol-chloroform-water to give 9a-bromo-16a-(nitromethyl) prednisone.

EXAMPLE 25 16a-(u-nitr0pr0pyl) cortisone A. ISa-(a-NITROPROPYL) PREGNANE3a-OL-11,20-

DIONE In the manner described in Example 1A, 16-pregnene- 3a-01-11,20-di0ne 3-acetate is reacted with l-nitropropane to give 160L-(ot-Dltl'Op10Py1) pregnane-3a-ol-1L20- dione.

IB. 16a-(a-NITROPROPYL) PREGNANE-3a,17a-DIOL- 11,20-DIONE The l6u-(a-nitropropyl) pregnane of above Example 25B is first brominated and then reacted with dimethylformamide and sodium acetate in the manner described in Example 10 to give 16a-(a-nitropropyl) pregnanc- 3a,17a,21-triol-11,20-dione 2l-acetate.

D. IGu-(a-NITROPROPYL) PREGNANE-17a,21-DIOL-3,

11,20-TRIONE 21-ACETATE The l6a-(oc-nitropropyl) pregnane-triol of above Example 25C is reacted with N-bromosuccinimide in the man ner of Example 1D to give 16a-(a-nitropropyl) pregnancl7a,2l-diol-3,11,20-trione 2l-acetate.

E. 4fl-BROMO-16a-(a-NITROPROPYL) PREGNANE-lia,

21-DIOL-3,11,20-TRIONE 21-ACETATE The 16a-(u-nitropropyl)-pregnane of above Example 25D is brominated in the manner described in Example IE to give 4/i-bromo-l6a-(a-nitropropyl) pregnane- 17a,21-dlO1-3,11,20-i1i0l16 21-acetate.

F. 16a-(a-NITROPROPYL) CORTISONE ZI-ACETATE The 4/3-bromopregnane of above Example 25B is first reacted with semicarbazide and thence with 80% acetic acid-water and 70% pyruvic acid in the manner described in Example IE to give l6a-(a-nitropropyl) cortisone Zl-acetate.

G. lfia-(a-NITROPROPYL) CORTISONE The 2l-acetate of above Example 25F is hydrolyzed with potassium bicarbonate in methanol-Water in the manner described in Example 2 to give 16ot-(a-nitropropyl) cortisone.

EXAMPLE 26 16a-(nitromethyl) -prednisne 21 acetate A. 16a- (NITROMETHYL) -ALLOPREGNANE-3B-OL11,20-

DIONE 3-acetate in 50 ml. of t-butyl alcohol containing about 1 g. of potassium t-butylateis reacted with 3 ml. of nitromethane in the manner described in Example 1A. The resultant product is isolated and purified in the described manner to give l6u-(nitro-methyl)-'allopregnane-3,B-ol- 11,20-dione. V

B. IGa-(NITROMETHYL)-ALLOPREGNANE-3fi,-l7a- DIOL-11,20-DIONE 16a(Nitromethy1) allopregnane 3 8 ol-ll,20-dio-ne, prepared as in Example 26A is reacted with p-toluene sulfonic acid in acetic anhydride in the manner of Example LB. The resultant product is isolated and purified to give 16cc (nitromethyl) -=allopregnane-3 3,l7a-dio l-l1,20- dione.

C. 16a- (NITROMETHYL) -ALLOPREG-NAN E-3,B,17 (1,21-

TRIOL-1L20-DIONE ZI-ACETATE In the manner of Example 1C, l6oz'(nitromethyl)alllopregnane-3 8,l7udiol-l1,20-dione, prepared as in Example 26B is brominated with bromine in chloroform and then treated with dimethyl formamidev The resultant product is isolated and purified in the described manner to give l6a (nitromethyl)-allopregnane-3fi,l7ut,21-triol- 11,20-dione ZI-acetate.

1). 16a- NITROMETHYL) -ALLOPREGNANE417 41,21- DIOL-3,11,20-TRIONE 21-ACETATE l6a-(Nitromethyl) allopregnane-SB,l7cc,2l-triol-3,20- dione 2l-acetate, the product of Example 26C, is reacted with N-bromosuccin-imide in the manner described in Example 1]) and the resultant product isolated and purified to give lfia-(nitromethyl)-a-llopregnane-l7a,2ldiol-3,l l,20-trione ZI-acetate.

E. 2,4-DI-BROMO-16a-(NITROMETHYL)-ALLOPREGNANE- 17a,21-DIOL-3,11,20:TRIONE 21ACETATE l6nw(Nitromethyl) allopregnane 17a,2l-diO1-3,l1,20 trione 2l-acet-ate, the compound of Example 26D, is dibrominated in the manner of Example 11A and the resultant product is isolated yielding 'a product substantially of 2,4-dibromo locdnitromethyl)-allopregnane-l7a,2ldiol3,ll,20-trione 21-acetate, of sutficient purity to. be used in the conversion described in the following procedure.

F. 16a-(NITROl\dETHYL) -PREDNISONE 21-ACETATE The 2,4-dibromide prepared as in above Example 26E is dehydrobrominated by means of dimethyl formamide in the manner of Example 11B. The resultant product is isolated and purified in the described manner to give l6a-(nitromethyD-prednisone ZI-acetate Am 237 my (6 14,600)

EXAMPLE 27 1611- (nitromethyl) -cortisone- 21 -acetate A. 2-IODO-16a-(NITROMETHYL) -CORTISONE 'ZI-ACETAIE A solution of 1.0 g. of 2,4-dibromo-16a-(nitromethyl)'- allopregnane-l7a,2l-diol-3,11,20-trione ZI-acetate, prepared as in Example 26E, in 50 of acetone containing 2.12 g. of sodium iodide is refluxed under nitrogen for 2 /2 hours. Excess of 0.1 N sodium thiosulfate solution is then added toremove iodine, then water is added and the mixture extracted with methylene chloride. The organic extracts are combined and evaporated to give a residue substantially of 2-iodo16a-(nitromethyl)-cortisone acetate which is'used without further purification in the reaction immediately following.

added and the mixture extracted with methylene chloride. 7

The organic extracts are washed with water, dried and evaporated to a residue. Crystallization of this residue from acetone-hexane gives 16a- (nitromethyl) -cortisone acetate.

EXAMPLE 28 16a-(u-nitr0butyl) cortisone A. lfia-(a-NITROBUTYL) PREGNANE-3a-OL-1L20- DIONE In the manner described in Example 1A, 16-preguene- 3a-ol-ll,20-dione 3-acetate is reacted with l-nitrobutane to give 16m-(a-nitrobutyl) pregnane-3a-o1-11,20-dione.

B. 1oa(a-NITROBUTYL) PREGNANEMJ'Ta-DIOL- 11,20-DIONE The 16a-(e-nitrobutyl) pregnane of above Example 28A is treated in the manner described in Example 13, and the resulting product isolated and purified in the described manner to give 16u-(a-nitrobutyl) pregnanc- 30:,17a-di01-1L20di011fi.

.c. IGa-(a-NITROBUTYL) PREGNANE-3a,17e,Zl-TRIOL- 11,20-DIONE ill-ACETATE The la-(a-nitrobutyl) pregnane of above Example 288 is first brominated and then reacted with dimethylformamide and sodium acetate in the manner described in Example lC to give l6a-(a-nitrobutyl) pregnanc- 3oz,l7a,2l-iI'iOl-11,20-di0116 ZI-acetate.

D. 16a-(a-NITROBUTYL) PREGNANE-17a,21-DIOL- 3,11,20-TRIONE Zl-ACETATE The 16a-(a-nitrobutyl) pregnane-triol of above Example 28C is reacted with N-bromosuccinimide in the manner of Example 1D to give l6ot-(a-nitrobuty1) pregnanc- 17a,21-diO1-3,1 1,20-trione ZI-acetate.

n. 4B-BROMO16a-(a-NITROBUTYL) PREGNANE- 17a,2l-DIOL-3,1l,ZO-TRIONE 21-ACETATE The 16m-(a-nitrobutyl) pregnane of above Example 28D is brominated in the manner described in Example 1E to give 4fl-bromo-16a-(a-nitrobutyl) pregnancl7a .2l-diol-3,11,20-trione 2'1-acetate.

F. lfia-(a-NITROBUTYL) CORTISONE ZI-ACETATE The 4fl-bromopregnane of above Example 28E is first reacted with semicarbazide and thence with 80% acetic acid-water and 70% pyruvic acid in the manner described in Example 1F to give 16a-(a-nitrobutyl) cortisone ZI-acetate.

G. lfia-(a-NITROBUTYL) CORTISONE The ZI-acetate of above Example 28F is hydrolyzed with potassium bicarbonate in methanol-water in the manner described in Example 2 to give 16rx-(a'nitr0- butyl) cortisone.

EXAMPLE 29 9a-flu0r0-16a- (nitromezhyl) cortisone 21 -acetate To a solution of 0.3 g. of the 21-acetate of 9a-fiuoro- 16a-(nitromethyl) hydrocortisone prepared as described in Example 7 in 15 ml. of acetone is added dropwise a solution of 60 mg. of chromium trioxide in 1 m1. of water and 3 ml. of acetic acid. The resulting mixture is allowed to stand for a period of about hours, then diluted with water and extracted with methylene chloride. The organic extracts are washed with water, dried over magnesium sulfate, filtered and evaporated in a residue which is crystallized from methanol to give 0.1 g. of 9a-fll1010 IGOL-(DjtfOlIlfithYl) cortisone ZI-acetate.

EXAMPLE 30 9a-fluar0-I6e-(nitromethyl) cortisone CHnOR '"OH Z wherein [W W is a member selected from the group consisting of -CH CH and CH=CH-; Y is a member selected from the group consisting of O, (H,ecOH) and (H,;3OH); X is a member selected from the group consisting of H and a halogen atom, said halogen atom having an atomic weight of less than 125, and X is H when Y is (H,o:OH); each of Z and Z is a member selected from the group consisting of H and alkyl radicals; and R is a member selected from the group consisting of H and acyl.

2. Compounds of the following general formula:

wherein X is a halogen of atomic weight less than 125. 5. The 21-lower alkanoates of the compounds of claim 4.

6. Compounds of the following general formula:

CIIgOl-I wherein each of Z and Z is lower alkyl.

7. The 21-lower alkanoates of the compounds of claim 6.

8. Compounds of the following general formula: 14. 9u-fiuoro-16u-(nitromethy1) prednisone.

15. 9a-fluo1'o-loa-(nitromethyl) prednisone ZI-acetate.

35203 16. 9m-fluoro-16u-(nitromethyl) prednisolone. I 17. 9u-fluoro-16m-(nitromethy1) prednisolone ZI-aco- L: z 5 tate. OHM if 18. lfiu-(nitromethyl) cortisone.

19. 16a-(nitromethy1) cortisone ZI-acetate. Hm N0: Z 20. 16a-(nitromethy1) hydrocortisone.

21. lfiu-(nitromethyl) hydrocortisone ZI-acetate. 10 22. 16a-(nitromethyl) prednisone. 23. 16a-(nitromethy1) prednisone ZI-acetate.

24. lfia-(nitromethyl) prednisolone.

wherein each of Z and Z1 is lower alkyL 25. IGa-(nitromethyl) prednisolone 21-aoetate.

9. The 21-lower alkanoates of the compounds of 16 References Cited in the file of this Patent claim 8.

10. 9a-fluo1'o-16a-(nitromethyl) cortisone. UNITED STATES PATENTS 11. 9a-fluoro-16a-(nitromethy1) cortisone 21-acetate. 2,671,084 Lincoln t a1 Mar. 2, 1954 12. 9a-fluoro-16a-(nitromethy1) hydrocortisone. 2,697,109 Dodson Dec. 14, 1954 13. 9a-fluoro-16u-(nitromethyl) hydrocortisone 21- 20 2,794, 15 D dson June 4, 1957 acetate. 

1. A 16A-(A-NITROALKYL)-PREGNENE HAVING THE FORMULA: 